Top Tweets for #fcuk3D

NANOLUC, BRET, QUANTUM DOTS and the RED-SHIFT!!! 📃 Semisynthetic, multicolor probes for bioluminescence imaging NanoLuc is one of the brightest and most widely used luciferase enzymes, however, the native blue emission of NanoLuc limits its applications due to poor sample penetration. NanoBiT, a split luciferase reporter derived from NanoLuc, consists of two subunits: LgBiT and HiBiT. Herein, we take advantage of bioluminescence resonance energy transfer (BRET) to RED-SHIFT the emission of NanoBiT by chemically labeling the N-terminus of HiBiT with fluorescent probes. Upon association of HiBiT and LgBiT, the appended fluorophores emit RED-SHIFTED LIGHT. When LgBiT is genetically encoded on the surface of mammalian cells, each probe produces unique emission profiles that can be distinguished by spectral phasor analysis microscopy. Moreover, by conjugating HiBiT peptides to fluorescent quantum dots, we extended BRET emission into the near-infrared. This modular system allows easy synthesis, rapid color switching, and tunable emission across the visible to near-infrared spectrum, providing a versatile platform for multicolor bioluminescence imaging across biological scales. https://t.co/JwXgpianKg 📃 An Engineered Amber-Emitting Nano Luciferase and Its Use for Immunobioluminescence Imaging in Vivo Here, we present the development of an NLuc mutant, QLuc, which catalyzes the oxidation of a synthetic QTZ luciferin for bright and RED-SHIFTED emission peaking at ∼ 585 nm. This amber-light-emitting luciferase-luciferin pair exhibited improved performance for imaging deep-tissue targets in live mice. Leveraging this novel bioluminescent reporter, we further pursued in vivo immunobioluminescence imaging (immunoBLI), which used a fusion protein of a single-chain variable antibody fragment (scFv) and QLuc for molecular imaging of tumor-associated antigens in a xenograft mouse model. As one of the MOST RED-SHIFTED NLuc variants, we expect QLuc to find broad applications in NONINVASIVE IMAGING IN MAMMALS. The activity of QLuc is independent of ATP, and thus, it remains functional in the extracellular space where ATP concentrations are low. Because of its bright and red-shifted emission and excellent photon penetration in mammalian tissue, we envisaged that QLuc could be used to tag antibodies or antibody fragments for imaging specific cell-surface markers in vivo. https://t.co/U62Dor2QEN PROMEGA CORP Functionalized nanoluc inhibitors EP3562823A1 [0092] If the luminescent enzyme is one that occurs naturally or is a recombinant or mutant luminescent enzyme, e.g. one which retains activity in a luciferase-coelenterazine or luciferase-luciferin reaction of a naturally occurring luminescent enzyme, it can be obtained readily from a culture of bacteria, yeast, mammalian cells, insect cells, plant cells, or the like, transformed to express a nucleic acid encoding the luminescent enzyme. Further, the recombinant or mutant luminescent enzyme can be derived from an in vitro cell-free system using a nucleic acid encoding the luciferase. https://t.co/Uu5YAhzPnO